Reconstitution of Functional Integrin αIIbβ3 and Its Activation in Plasma Membrane-Mimetic Lipid Environments
The study of the platelet receptor integrin αIIbβ3 in a membrane-mimetic environment without interfering signalling pathways is crucial to understand protein structure and dynamics. Our understanding of this receptor and its sequential activation steps has been tremendously progressing using structu...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-06-01
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| Series: | Membranes |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2077-0375/11/7/499 |
| _version_ | 1797528208597319680 |
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| author | Una Janke Alexandra Mitlehner Aileen Weide Theresia Gutmann Mihaela Delcea |
| author_facet | Una Janke Alexandra Mitlehner Aileen Weide Theresia Gutmann Mihaela Delcea |
| author_sort | Una Janke |
| collection | DOAJ |
| description | The study of the platelet receptor integrin αIIbβ3 in a membrane-mimetic environment without interfering signalling pathways is crucial to understand protein structure and dynamics. Our understanding of this receptor and its sequential activation steps has been tremendously progressing using structural and reconstitution approaches in model membranes, such as liposomes or supported-lipid bilayers. For most αIIbβ3 reconstitution approaches, saturated short-chain lipids have been used, which is not reflecting the native platelet cell membrane composition. We report here on the reconstitution of label-free full-length αIIbβ3 in liposomes containing cholesterol, sphingomyelin, and unsaturated phosphatidylcholine mimicking the plasma membrane that formed supported-lipid bilayers for quartz-crystal microbalance with dissipation (QCM-D) experiments. We demonstrate the relevance of the lipid environment and its resulting physicochemical properties on integrin reconstitution efficiency and its conformational dynamics. We present here an approach to investigate αIIbβ3 in a biomimetic membrane system as a useful platform do dissect disease-relevant integrin mutations and effects on ligand binding in a lipid-specific context, which might be applicable for drug screening. |
| first_indexed | 2024-03-10T09:54:54Z |
| format | Article |
| id | doaj.art-930be4f5577846f6a2d0b5d891691a9a |
| institution | Directory Open Access Journal |
| issn | 2077-0375 |
| language | English |
| last_indexed | 2024-03-10T09:54:54Z |
| publishDate | 2021-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Membranes |
| spelling | doaj.art-930be4f5577846f6a2d0b5d891691a9a2023-11-22T02:27:59ZengMDPI AGMembranes2077-03752021-06-0111749910.3390/membranes11070499Reconstitution of Functional Integrin αIIbβ3 and Its Activation in Plasma Membrane-Mimetic Lipid EnvironmentsUna Janke0Alexandra Mitlehner1Aileen Weide2Theresia Gutmann3Mihaela Delcea4Institute of Biochemistry, University of Greifswald, Felix-Hausdorff-Straße 4, 17489 Greifswald, GermanyInstitute of Biochemistry, University of Greifswald, Felix-Hausdorff-Straße 4, 17489 Greifswald, GermanyInstitute of Biochemistry, University of Greifswald, Felix-Hausdorff-Straße 4, 17489 Greifswald, GermanyPaul Langerhans Institute Dresden of the Helmholtz Zentrum München at the University Hospital and Faculty of Medicine Carl Gustav Carus of TU Dresden, 01307 Dresden, GermanyInstitute of Biochemistry, University of Greifswald, Felix-Hausdorff-Straße 4, 17489 Greifswald, GermanyThe study of the platelet receptor integrin αIIbβ3 in a membrane-mimetic environment without interfering signalling pathways is crucial to understand protein structure and dynamics. Our understanding of this receptor and its sequential activation steps has been tremendously progressing using structural and reconstitution approaches in model membranes, such as liposomes or supported-lipid bilayers. For most αIIbβ3 reconstitution approaches, saturated short-chain lipids have been used, which is not reflecting the native platelet cell membrane composition. We report here on the reconstitution of label-free full-length αIIbβ3 in liposomes containing cholesterol, sphingomyelin, and unsaturated phosphatidylcholine mimicking the plasma membrane that formed supported-lipid bilayers for quartz-crystal microbalance with dissipation (QCM-D) experiments. We demonstrate the relevance of the lipid environment and its resulting physicochemical properties on integrin reconstitution efficiency and its conformational dynamics. We present here an approach to investigate αIIbβ3 in a biomimetic membrane system as a useful platform do dissect disease-relevant integrin mutations and effects on ligand binding in a lipid-specific context, which might be applicable for drug screening.https://www.mdpi.com/2077-0375/11/7/499liposomeslipidsintegrin αIIbβ3reconstitutiondetergents |
| spellingShingle | Una Janke Alexandra Mitlehner Aileen Weide Theresia Gutmann Mihaela Delcea Reconstitution of Functional Integrin αIIbβ3 and Its Activation in Plasma Membrane-Mimetic Lipid Environments Membranes liposomes lipids integrin αIIbβ3 reconstitution detergents |
| title | Reconstitution of Functional Integrin αIIbβ3 and Its Activation in Plasma Membrane-Mimetic Lipid Environments |
| title_full | Reconstitution of Functional Integrin αIIbβ3 and Its Activation in Plasma Membrane-Mimetic Lipid Environments |
| title_fullStr | Reconstitution of Functional Integrin αIIbβ3 and Its Activation in Plasma Membrane-Mimetic Lipid Environments |
| title_full_unstemmed | Reconstitution of Functional Integrin αIIbβ3 and Its Activation in Plasma Membrane-Mimetic Lipid Environments |
| title_short | Reconstitution of Functional Integrin αIIbβ3 and Its Activation in Plasma Membrane-Mimetic Lipid Environments |
| title_sort | reconstitution of functional integrin αiibβ3 and its activation in plasma membrane mimetic lipid environments |
| topic | liposomes lipids integrin αIIbβ3 reconstitution detergents |
| url | https://www.mdpi.com/2077-0375/11/7/499 |
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